Whole-cell patch-clamp experiments were performed with HEK293 cells heterologously expressing human NaV1.7 (SB Drug Discovery) using a QPatch 16X automated electrophysiology platform (Sophion Bioscience). The extracellular solution contained the following: 70 mM NaCl, 70 mM choline chloride, 4 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 10 mM Hepes, and 10 mM glucose (pH 7.4; osmolarity, 305 mosmol). The intracellular solution contained the following: 140 mM CsF, 1 mM:5 mM EGTA/CsOH, 10 mM Hepes, and 10 mM NaCl (pH 7.3 with CsOH; osmolarity, 320 mosmol). MIITX1-Mg1a was diluted in extracellular solution with 0.05% BSA at a concentration of 3 μM. MIITX1-Mg1a effects were compared to pretoxin control conditions on the same cell with incubation times for buffer and MIITX1-Mg1a of 5 min each. NaV1.7 currents were induced with a holding potential of −90 mV followed by a 50-ms test pulse to −20 mV and a 0.05-Hz pulse frequency.

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